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      Establishment of a panel of in‐house polyclonal antibodies for the diagnosis of enterovirus infections

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          Abstract

          The aim of this study was to establish a reliable method of virus detection for the diagnosis of critical enterovirus infections such as acute infective encephalitis, encephalomyelitis and myocarditis. Because histopathological and immunohistochemical analyses of paraffin‐embedded tissues play an important role in recognizing infectious agents in tissue samples, six in‐house polyclonal antibodies raised against three representative enteroviruses using an indirect immunofluorescence assay and immunohistochemistry were examined. This panel of polyclonal antibodies recognized three serotypes of enterovirus. Two of the polyclonal antibodies were raised against denatured virus particles from enterovirus A71, one was raised against the recombinant VP1 protein of coxsackievirus B3, and the other for poliovirus type 1 were raised against denatured virus particles, the recombinant VP1 protein and peptide 2C. Western blot analysis revealed that each of these antibodies recognized the corresponding viral antigen and none cross‐reacted with non‐enteroviruses within the family Picornaviridae. However, all cross‐reacted to some extent with the antigens derived from other serotypes of enterovirus. Indirect immunofluorescence assay and immunohistochemistry revealed that the virus capsid and non‐structural proteins were localized in the cytoplasm of affected culture cells, and skeletal muscles and neurons in neonatal mice experimentally‐infected with human enterovirus. The antibodies also recognized antigens derived from recent clinical isolates of enterovirus A71, coxsackievirus B3 and poliovirus. In addition, immunohistochemistry revealed that representative antibodies tested showed the same recognition pattern according to each serotype. Thus, the panel of in‐house anti‐enterovirus polyclonal antibodies described herein will be an important tool for the screening and pathological diagnosis for enterovirus infections, and may be useful for the classification of different enterovirus serotypes, including coxsackieviruses A and B, echoviruses, enterovirus A71 and poliovirus.

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          An epidemic of enterovirus 71 infection in Taiwan. Taiwan Enterovirus Epidemic Working Group.

          Monto Ho, Eng-Rin Chen, Kwo-Hsiung Hsu … (1999)
          Enteroviruses can cause outbreaks of hand-foot-and-mouth disease (characterized by vesicular lesions on the hands, feet, and oral mucosa) or herpangina, usually without life-threatening manifestations. In 1998 an epidemic of enterovirus 71 infection caused hand-foot-and-mouth disease and herpangina in thousands of people in Taiwan, some of whom died. We assessed the epidemiologic aspects of this outbreak. Cases of hand-foot-and-mouth disease or herpangina in ambulatory patients were reported to the Taiwan Department of Health by a mean of 818 sentinel physicians. Severe cases in hospitalized patients were reported by 40 medical centers and regional hospitals. Viruses were isolated by 10 hospital laboratories and the department of health. The sentinel physicians reported 129,106 cases of hand-foot-and-mouth disease or herpangina in two waves of the epidemic, which probably represents less than 10 percent of the estimated total number of cases. There were 405 patients with severe disease, most of whom were five years old or younger; severe disease was seen in all regions of the island. Complications included encephalitis, aseptic meningitis, pulmonary edema or hemorrhage, acute flaccid paralysis, and myocarditis. Seventy-eight patients died, 71 of whom (91 percent) were five years of age or younger. Of the patients who died, 65 (83 percent) had pulmonary edema or pulmonary hemorrhage. Among patients from whom a virus was isolated, enterovirus 71 was present in 48.7 percent of outpatients with uncomplicated hand-foot-and-mouth disease or herpangina, 75 percent of hospitalized patients who survived, and 92 percent of patients who died. Although several enteroviruses were circulating in Taiwan during the 1998 epidemic, enterovirus 71 infection was associated with most of the serious clinical manifestations and with nearly all the deaths. Most of those who died were young, and the majority died of pulmonary edema and pulmonary hemorrhage.
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            Deaths of children during an outbreak of hand, foot, and mouth disease in sarawak, malaysia: clinical and pathological characteristics of the disease. For the Outbreak Study Group.

            L G Chan, U. Parashar, M S Lye … (2000)
            From April through June 1997, 29 previously healthy children aged <6 years (median, 1.5 years) in Sarawak, Malaysia, died of rapidly progressive cardiorespiratory failure during an outbreak of hand, foot, and mouth disease caused primarily by enterovirus 71 (EV71). The case children were hospitalized after a short illness (median duration, 2 days) that usually included fever (in 100% of case children), oral ulcers (66%), and extremity rashes (62%). The illness rapidly progressed to include seizures (28%), flaccid limb weakness (17%), or cardiopulmonary symptoms (of 24 children, 17 had chest radiographs showing pulmonary edema, and 24 had echocardiograms showing left ventricular dysfunction), resulting in cardiopulmonary arrest soon after hospitalization (median time, 9 h). Cardiac tissue from 10 patients showed normal myocardium, but central nervous system tissue from 5 patients showed inflammatory changes. Brain-stem specimens from 2 patients were available, and both specimens showed extensive neuronal degeneration, inflammation, and necrosis, suggesting that a central nervous system infection was responsible for the disease, with the cardiopulmonary dysfunction being neurogenic in origin. EV71 and possibly an adenovirus, other enteroviruses, or unknown cofactors are likely responsible for this rapidly fatal disease.
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              An outbreak of hand, foot, and mouth disease associated with subgenotype C4 of human enterovirus 71 in Shandong, China.

              Yong Zhang, Xiao-Juan Tan, Hai-Yan Wang … (2009)
              An outbreak of hand, foot, and mouth disease (HFMD) included 1149 people in Linyi City, Shandong Province, China, in 2007: three children died. To characterize the pathogens responsible for this outbreak and to analyze their genetic features. A total of 233 clinical specimens were collected from 105 hospitalized patients, including 11 patients with severe HFMD. Virological investigations (direct RT-PCR, viral isolation and molecular identification) and phylogenetic analysis were performed. Human enterovirus 71 (HEV71) was the main pathogen that caused this outbreak, based on clinical manifestations, epidemiological data, and laboratory results. Phylogenetic analysis indicated that the Shandong HEV71 isolates belonged to 3 lineages in subgenotype C4. Subgenotype C4 could be further divided into two clusters (C4a and C4b), which corresponded to two time periods. Cluster C4a HEV71 has been the predominant virus circulating in mainland China in the past 5 years. The 2007 HFMD outbreak was mainly caused by HEV71 subgenotype C4 with 3 transmission chains. This virus has been continuously circulating in China since 1998. The Shandong strains co-evolved with isolates from other provinces in mainland China and neighboring countries.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Neuropathology
                Journal ID (iso-abbrev): Neuropathology
                Journal ID (doi): 10.1111/(ISSN)1440-1789
                Journal ID (publisher-id): NEUP
                Title: Neuropathology
                Publisher: John Wiley and Sons Inc. (Hoboken )
                ISSN (Print): 0919-6544
                ISSN (Electronic): 1440-1789
                Publication date (Electronic): 28 September 2014
                Publication date (Print): April 2015
                Volume: 35
                Issue: 2 ( doiID: 10.1111/neup.2015.35.issue-2 )
                Pages: 107-121
                Affiliations
                [ 1 ] Department of Pathology National Institute of Infectious Diseases Aichi Japan
                [ 2 ] Department of Virology II National Institute of Infectious Diseases Aichi Japan
                [ 3 ] Department of Virology and Viral Infections Faculty of Veterinary Medicine Nippon Veterinary and Life Science University Aichi Japan
                [ 4 ] Neurovirology Project Tokyo Metropolitan Institute of Medical Science Aichi Japan
                [ 5 ] Department of Microbiology and Medical Zoology Aichi Prefectural Institute of Public Health Aichi Japan
                Author notes
                [*] [* ]Correspondence: Noriyo Nagata, DVM, PhD, 4‐7‐1 Gakuen, Musashimurayama, Tokyo 208‐0011, Japan. Email: nnagata@ 123456niid.go.jp
                Article
                Publisher ID: NEUP12171
                DOI: 10.1111/neup.12171
                PMC ID: 7168124
                PubMed ID: 25263613
                SO-VID: 13f5a7a0-d06a-46f0-b66b-3f8e6f6d7414
                Copyright © © 2014 Japanese Society of Neuropathology
                License:

                This article is being made freely available through PubMed Central as part of the COVID-19 public health emergency response. It can be used for unrestricted research re-use and analysis in any form or by any means with acknowledgement of the original source, for the duration of the public health emergency.

                History
                Date received : 24 August 2014
                Date accepted : 02 September 2014
                Page count
                Pages: 15
                Funding
                Funded by: Ministry of Education, Culture, Sports, Science, and Technology, Japan
                Award ID: 23590554
                Funded by: Ministry of Health, Labor, and Welfare, Japan
                Award ID: H23‐Shinko‐Ippan‐007
                Award ID: H25‐Shinko‐Ippan‐012
                Categories
                Subject: Original Articles
                Subject: Original Article
                Custom metadata
                source-schema-version-number 2.0
                cover-date April 2015
                details-of-publishers-convertor Converter:WILEY_ML3GV2_TO_JATSPMC version:5.8.0 mode:remove_FC converted:15.04.2020

                Keywords: enterovirus,immunohistochemistry,paraffin‐embedded tissue,polyclonal antibody,viral encephalitis
                Data availability:
                Keywords: enterovirus, immunohistochemistry, paraffin‐embedded tissue, polyclonal antibody, viral encephalitis

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